Nitrogen containing steroids



3,409,641 NITROGEN CONTAINING STEROIDS Milton Heller and SeymourBernstein, New City, N.Y., assignors to American Cyanamid Company,Stamford, Conn., a corporation of Maine No Drawing. Filed May 16, 1966,Ser. No. 550,174

and lower alkanoyl CH3 o l:H

and R is selected from the group consisting of Claims. (Cl. 260397-4) /HThis invention relates to new steroid compounds. More N=N+=N7 N1wra]kylparticularly, it relates to 16,17-substituted pregnenes. 10 alkyl H Thenovel pregnenes of the present invention may be il- N N -NCNH1 lustratedby the following formula: \lower alkyl \lower alkamyl' I CH3 CH3 H CH3R1 OH; R Z-ocmm E-ocm-Q /H CgHg -N -N and R fi-CH and lower alkauoyl 0(I) (II) COOH wherein R is Selected from the group consisting of Thepresent compounds are substantially insoluble droxyr lower alkanoyloxyformyloxy and in water and soluble 1n the usual organic solvents suchas, for example, lower alkanols, ether, chloroform acetone, hexane, etc.R 1 t d f th f The compounds of the present invention are prepared 1 15Se cc 6 mm 6 group consls mg 0 by starting with3B-acetoxy-16a-hydroxypregn-S-en-ZO-one (3H3 which in turn is describedby Cole et al., J. Org. Chem. 19, 11 131 (1954). The following flowsheetshows the preparation of the present compounds FLOWSHEET CH; CH; 3H3

HOG-H HgC I "OSOQCHS A O- HO- (n c (In (In) CH3 CH CH3 HOCH HO-CH HCO CHI H3O I N Q I/ .AcO- (IV) HOCH H O NHCONH: l/ \J l H30 l/ 8 HO (IX) HaCIn the above fiowsheet 3fi-acetoxy-16a-hydroxypregn-5- en-20-one (I) isreacted with methanesulfonyl chloride absolute lower alkanol produces3fi-acetoxy-16a-methanesulfonyloxypregn-S-en-20,8-01 (III). When the2013-01 is treated with sodium azide in the presence of N-methyl-2-pyrrolidone and a lower alkyl alcohol, the product 36- acetoxy-l68-azidopregn-S-en-ZOfl-ol (IV) is obtained. When compound (IV) is heatedwith lithium aluminum hydride in a solvent, 163-aminopregn-5-en-3fi,20/8-diol (V) is obtained. When the lattercompound is recrystallized from acetone or acetone-petroleum ether,2,2,6'(R)- trimethyl 2'3,4',5' tetrahydro 1',3' oxazino [4,5:16 3,179]androst-5-en-3,8-ol (IX) is obtained. When the latter compound istreated with acetic anhydride in the presence of pyridine,16/3-acetamidopregn-5-ene- 3/8,20 3-diol diacetate (VIII) is obtained.Treating the trimethyl 1,3-oxazine (IX) with hydrogen chloride inmethylene chloride produces 16,8-aminopregn-5-ene-3fi, 20,8-diolhydrochloride (VI). Heating the N-acetyl 3,20-diacetate (VIII) withpotassium carbonate in the presence of a lower alkanol produces16,8-acetamidopregn-5-ene- 3,8,20/8-diol (XI). The latter compound whenheated with aluminum isopropoxide in a solvent, produces 165-acetamido-20 8-hydroxypregn-4-ene-3-one (XIV). The amino-diol (V) isheated with nitrourea in the presence of a lower alkanol to produce16;8carbamido producesN-carbobenzoxy-3,B,20,8-dihydroxypregn-5-en-16fi-yl-amine (X). The16,8-an1ino 3,20-diol (V) when heated with phthalic anhydride ofdioxane, produces l65-(o-carboxybenzamido)pregn-S- ene-3B,20,B-diol(XIII). Heating 16fi-aminopregn-5-ene- 3,3,20,6-idol (V) with formicacid and formaldehyde produces 3',6(R) dimethyl 2',3',4,5 tetrahydro1,3- oxazinol[4',5:16,6,l7fl]androst-5-en-3,B-ol (XVI) and also heatedwith formaldehyde in the presence of a lower alkanol,6(R)-methyl-2,3,4,5-tetrahydro-1',3-oxazino-[4,5':16/3,17fi]androst-5-ene-3B-ol (XIX) is produced. The lattercompound when reacted with acetic anhydride in pyridine, produces3'-acetyl-6'-(R)methyl-2'-3,4,5- tetrahydro 1,3' oxazino[4',5:l6,8,17,8] androst 5 en- Sfl-ol acetate (XX). The reaction of16,8-aminopregn-5- ene-3,8,20fi-diol (V) in pyridine with ethylchloroformate produces N-carbethoxy-35,20,8-dihydroxypregn-5-en-16B-yl-amine (XXII). The latter compound heated with lithium aluminumhydride in the presence of the tetrahydrofuran produces16fi-methylaminopregn-5-ene-33,2018- diol (XXIV). The N-acetyl diacetate(VIII) heated with lithium aluminum hydride in the presence oftetrahydrofuran gives 16p ethylaminopregn 5 I18-3fl,20y3d10l (XII). Whenthe N-methyl oxazine (XVI) is heated with lithium aluminum hydride intetrahydrofuran, the product 165 dimethylaminopregn 5 en 35,206 diol(XXIII) is obtained. Further when the N-acetyl oxazine (XX) is heatedwith lithium aluminum hydride in tetrahydrofuran, the product 168-ethylmethylaminopregn-S- ene-3fl,20fl-diol (XXI) is obtained. Stillfurther, when the trimethyl-1,3-oxazine (IX) is heated with lithiumaluminum hydride in a solvent, the product16,8-isopropylaminopregn-5-ene-3fl,20fi-diol(XVIII) is obtained. Whenthe 1613 ethylamine 3,8,20/3 diol (XII) is reacted with acetic anhydridein the presence of pyridine the productN-acetyl-16p-ethylaminopregn-5-ene-3/3,20,6-diol diacetate (XV) isobtained.

The compounds of the present invention possess antibacterial andanti-fungal properties and are therefore useful in the treatment ofconditions resulting from the presence of bacteria and fungi.

The following examples illustrate in detail the preparation ofrepresentative 16,17-substituted pregnenes of this invention.

Example 1.-Preparation of 38-acetoxy-l6umethanesulfonyloxypregn-S-en-ZO-one (II) solutioncontaining 1 g. of 3 3-acetoxy- [prepared according 113$; 1742 1710,1361, 1248 and 1182 cmf Example 2.Preparation of3fi-acetoxy-16amethanesulfonyloxypregn-S-en-20 3-01 (III) A mixture of5.2 g. of the methanesulfonate-ZO-one (product of Example 1) and 5.0 g.of sodium borohydride in absolute ethanol (780 ml.) is stirred at roomtemperature for 75 minutes, whereupon complete solution is effected. Thesolution is added to ice-water and the resultant precipitate (3.48 g.)crystallized from dilute methanol to giveSB-acetoxy-l6a-methanesulfonyloxypregn-5-en-20/3-ol, melting point158.5-159 C. Crystallization from a mixture of acetone-hexane gives ananalytical sample: melting point 161.5-162" C.; (chloroform);

11533502 1739, 1342, 1260 and 1178.

Example 3.-Preparation of 3fi-acetoxy-16 8- azidopregn-5-en-20 3-ol (IV)A mixture of the methanesulfonate compound, (III), (0.57 g.) (product ofExample 2) and sodium azide (0.45 g.) in 23.75 ml. ofN-methyl-Z-pyrrolidone and 1.25 ml. of t-butyl alcohol is heated on thesteam bath 1 533580 2150, 1738 and 1260 cmr Example 4.Preparation of16B-aminopregn- 5-ene-3fl,20 3-diol (V) A slurry of the16,8-azido-compound, (IV), (3.73 g.) (product of Example 3) and lithiumaluminum hydride (5 .0 g.) in 200 ml. of ether is refluxed for 16 hours.The excess hydride is cautiously reacted with a saturated solution ofpotassium and sodium tartrate. The mixture is filtered and the residueboiled in ethyl acetate. When the ethyl acetate filtrate from theresidue is combined with the above ether filtrate, a precipitate, (V),(1.475 g.) appears and is collected, melting point 238-242 C. Ananalytical sample is crystallized from ethyl acetate, melting point241-242 C.; [04 5 1 (methanol);

#55; 3300 1605 and 1050 cm.

Example 5. Preparation of 2',2',6'(R) -trimethyl-2',3', 4,5' tetrahydro1',3-oxazino[4,5':1618,17,8]androst- 5-en-3fl-ol (IX) 7 sample isobtained by crystallization from acetone, melting point l76.5-178.5 C.;[011 -8 (methanol).

The same compound, (IX), is also obtained directly by crystallizationfrom the l6j8-amine (V) from acetone.

Example 6.-Preparation of 16fl-acetamidopregn-5- ene-3B,20;8-dioldiacetate (VIII) Example 7.-Preparation of 16B-aminopregn-5-ene- 35,20fi-diol hydrochloride (VI) Gaseous hydrogen chloride is added to asolution of the 1,3-oxazine, (IX), (0.22 g.) the product of Example 5,in 10 ml. of methylene chloride for 30 seconds at room temperature. Thesolvent is removed under reduced pressure and crystallization fromacetone gives a solid (0.135 g.), melting point about 210 C. Repeatedrecrystallization from methanol-ether gives (VI), melting point 331.5-332 C. (dec.)', [111 --37 (methanol);

1606 and 1050 cmr' Example 8.-Preparation of 16fl-acetamidopregn-5-ene-3B,20{3-diol (XI) To a solution of the N-acetyl 3,20-diacetate,(VIII), (2.3 g.) (product of Example 6) in 100 ml. methanol is added 20ml. of 10% potassium carbonate and the resultant solution is refluxed.Complete reaction does not take place until two hours elapses accordingto infrared spectrum determination. Pouring the refluxed solution into alarge excess of water and crystallizing from dilute methanol gives (XI),1.7 g., melting point 28l.5-282.5 C. The analytical sample melted at283-284 C; [(11 -27 (methanol).

Example 9.-Preparation of l6B-acetamido-2013- hydroxypregn-4-ene-3-one(XIV) A mixture of the N-acetyl-3B,20B-diol (XI), (0.6 g.) (product ofExample 8) in 150 ml. of toluene and 8 ml. of cyclohexanone is distilleduntil about 50 ml. of solvent is removed. An aluminum isopropoxidesolution (0.06 g./ml.) in 10 ml. of toluene is added and the mixture isrefluxed for 18 hours. A saturated solution of potassium and sodiumtartrate (50 ml.) is added to the refluxed solution and the resultantmixture stream distilled until all the cyclohexanone is removed. Thesolids (0.22 g.) are filtered. The residue is heated in acetone and theremaining residue (0.074 g.) which is starting material, is collected.The material in the filtrate (0.217 g.) is put on an activated magnesiumsilicate column (40 g.). Collection of the eluates from petroleum ether(boiling range 30-60 C.):acetone (1:1) yields 0.12 g. of a glass.Crystallization from a mixture of acetone and hexane gives 0.048 g. of(XIV): melting point 262-263 C.; [M +49 (methanol);

r3332 251 my. (613,900). Example l0.-Preparation of 16fi-carbamidopregn-5-ene-3p,20fi-diol (VII) A solution of the amino-diol, (V), (1.0 g.)(product of Example 4) and nitrourea (0.364 g.) in 20 ml. of 95% ethanolis refluxed 15 minutes whereupon a copious precipitate is formed. Thisprecipitate is collected to yield C. The infrared spectrum is identicalto 8 0.66 g. (VII) melting point 251-252" C.', [M -22 (methanol).Crystallization from methanol does not improve the melting point.

Example 11.-Preparation of N-carbobenzoxy-3B,ZOB-dihydrovypregn-S-en-l6B-yl-amine (X) To a solution of16/3-aminopregn-5-ene-3{3,20 8-di0l, (V), (1.0 g.) (product of Example4) in 20 ml. of pyridine is added 1.0 ml. of carbobenzoxy chloride at 0C. The mixture is allowed to stand at this temperature for one-half hourand then at room temperature for one hour. The mixture is then pouredinto ice-water and the resultant precipitate (0.14 g.) collected.Crystallization gives 0.21 g. of crude (X), melting point 148-156 C.which further purified as indicated by thin layer chromatography(T.L.C). A 70 mg. portion of this crude sample is placed on a T.L.C.plate (silica gel, 20 cm. x 20 cm. x 0.5 mm.) and developed for 2 hoursin the system benzenezacetonezwater (2:1:2) (upper phase). The bandcontaining the product (about 2.5 to 3.0 cm. from the origin) is elutedwith acetone and the product is crystallized as a solvated material:0.046 g.; melting point 164.5- 165.5 C.;

1 5?; 1505 and 1695 cm.

Example 12.-Preparation of 16B(o-carboxybenzamide)-pregn-5-ene-3B,20B-diol (XIII) The solvent is removed residuecrystallized from dilute methanol to give the hymelting point 188-190 C.

droscopic product: 0.35 g.; (bubbles). An additional 0.44 g. of productis collected as a post-precipitate: B11 -24.5 (methanol);

1 5?; 1709 and 1640 cm." Example l3.-Preparation of 3,6(R)-dimethyl-2',3,4, 5' tetrahydro 1',3-oxazino[4',5:163,173] androst-S-en-3i3-ol (XVI) and 3',6(R)-dimethyl-2',3,4',5-tetrahydro 1',3'oxazino[4',5:165,175]androst-5-en-3B-ol formate (XVII) A mixture of1618-aminopregn-5-ene-3,6,20[3-diol, (V), (2.125 g.) (product of Example4) formic acid (30 ml.) and 40% formaldehyde (30 ml.) is refluxed for 4hours and then poured into ice-water. The water solution is extractedwith ether, then made basic with concentrated potassium hydroxide. Theprecipitate is collected. Crystallization from acetone-water and arecrystallization from acetone-hexane afiords 0.41 g. of (XVI), meltingpoint 186-188 C.; [a] -33 C. (chloroform).

Crystallization of the mother liquors from methanolwater gives 0.196 g.of the formate ester, (XVII), melting point 156-159 C.; [@1 -55.5(chloroform).

Example 14.Preparation of 6(R)-methyl-2,3',3',4-tetrahydro-1,3-oxazino[4',5' 1613,175-androst-5-en-36- ol (XIX) Asolution of 16B-aminopregn-5-ene-3l3,20fi-diol (V) (0.5 g.) (product ofExample 4) in 40% formaldehyde (25 ml.) and methanol (25 ml.) isrefluxed for 1 /2 hours. The methanol is removed under reduced pressureand the precipitate which formed is collected. Crystallization from amixture of methylene chloride, methanol and ether gives 0.235 g. of(XIX), melting point 252-255 C. An analytical sample melts at 264266 C.

Example 15.--Preparation of 3-acetyl-6'(R)-methyl- 2,3,4',5' tetrahydro1',3' oxazino[4',5:16,l7;3] androst-S-en-Bfl-ol acetate (XX) A mixtureof 6'(R)-methyl-2,3',4,5-tetrahydro-1,3'-oxazino[4,5:16,8,1713]androst-5-ene-3 8-ol (XIX) (0.134 g.) (product ofExample 14) in chloroform (4 rnl.), pyridine (4 rnl.), and aceticanhydride (2 ml.) is heated on a steam bath until solution is effected,about 15 minmagnesium silicate column (25 g.). Elution with petroleumether (30-60 C.):acetone (43.2) gives a solid which crystallizes fromacetone-hexane to yield 0.10 g. of (XX), melting point 216219 C. Ananalytical sample melts at 224.5-226 C.; [a] 53.5 (chloroform).

Example 16.Preparation ofN-carbethoxy-3;3,20fidihydroxypregn-S-en-16fl-yl-amine (XXIII) To an icecold solution of IGfi-aminopregn-S-ene- 3,8,20,8-diol (V) (1.0 g.) theproduct of Example 4, in

for one hour. The mixture is poured into ice-water and the resultantprecipitate is collected and crystallized from dilute methanol to give0.955 g. of (V), melting point 191-195 C. An analytical sample iscrystallized from acetone-hexane and melts at 197198.5 C.; [(21 35(chloroform).

Example 17.Preparation of 16,8-methylaminopregn-5- ene-3B,20fi-diol(XXIV) A mixture of the N-carbethoxy-diol (XXII) (0.533 g.) 16) andlithium aluminum hydride (0.5 g.) in 50 ml. tetrahydrofuran is stirredand refluxed from methanol-water, 0.43 g. (XXIV), melting point194.5-198" C. An analytical sample crystallized from aectone-hexanemelts at 216- 217 C.; [111 17 (methanol).

Example 18.Preparation of 16,3-ethylaminopregn-5- ene-3 3,20fl-diol(XII) from dilute acetone followed by crystallization fromacetone-hexane yields 0.72 g. of (XII), melting point 172.5173 C. Ananalytical sample melts at 181.5- 182.5 C.; [aJ -l7 (methanol).

Example 19.Preparation of l6 3-dimethylamin0pregn-5- ene-3 3,20,B-diol(XXIII) g. of (XXIII), melting point 168-l73 C. An analytical sample,crystallized from acetone-hexane melts at 180.5-182 C.; [a] 23"(chloroform).

Example 20.Preparation of 16fl-ethylmethylaminopregn- 5-ene-3/3,203-diol (XXI) A mixture of the N-acetyl oxazine (XX) (1.242 g.) (productof Example 15) and lithium aluminum hydride (2.0 g.) in tetrahydrofuranis stirred and refluxed for 40 worked up as in the preparation of the16-ethylarnino compound (XII) (product of Example 18), to give, aftercrystallization from dilute methanol, 0.95 g. of (XXI), melting pointabout 178- 183 C. The analytical sample, crystallized fromacetonehexane, melts at 178180 C.; [a];;, 39 (chloroform).

10 Example 21. Pr'eparation of IGB-isopropylaminepregn- 5-ene-38,20,8-diol (XVIII) A mixture of the trimethyl-L3-oxazine (IX) (0.215g.) (product of Example 5) 'Qystallization of the remaining residuegives 0.12 g. (XVIII), melting point about C. An analytical sample meltsat 169.5- 170.5 C.; [a] 24.5 (methanol).

Example 22.-Preparation ofN-acetyl-16fl-ethylaminopregn-5-cne-3fi,20fl-diol diacetate (XV) CH3 OH:

R CH3 wherein R is selected from the group consisting of hydroxy, loweralkanoyloxy, formyloxy and R: Ra

and

o CHaCHI o (i-@- R is selected from the group consisting of 6" H0t 3Hand lower and R is selected from the group consisting of -N=N+= NH -NH.CHl,

loweralkyl H lower alkanoyl N-lower alkyl, -N

lower alkyl 8. A pregnene according to claim 1, 16p-dimethylaminopregn-S-cnc-3l3,20;3-di0l.

9. A pregnene according to claim 1, 16B-methylaminopregn-5-ene-3fi,20;8-diol.

10. A pregncnc according to claim 1,16B-cthy1rnethylaminopregn-S-cne-313,20B-diol.

References Cited UNITED STATES PATENTS 2/1966 Bergstrom.

H. A. FRENCH, Primary Examiner.

1. A PREGNENE SELECTED FROM THE GROUP CONSISTING OF THE FORMULAS: